肝脏
肝髒
간장
CHINESE HEPATOLOGY
2014年
5期
311-315
,共5页
张晓培%秦爱兰%邓长卿%甘建和%石翠翠%刘涛
張曉培%秦愛蘭%鄧長卿%甘建和%石翠翠%劉濤
장효배%진애란%산장경%감건화%석취취%류도
脐带间充质干细胞%原代肝细胞%肝卵圆细胞%增殖%白蛋白
臍帶間充質榦細胞%原代肝細胞%肝卵圓細胞%增殖%白蛋白
제대간충질간세포%원대간세포%간란원세포%증식%백단백
Umbilical cord mesenchymal stem cells%Primary hepatocyte%Hepatic oval cells%Proliferation%Albumin
目的:探讨大鼠脐带间充质干细胞(UMSC)对原代大鼠肝细胞及肝卵圆细胞增殖与功能的影响。方法采用2-乙酰氨基芴加肝脏三分之二切除术建立肝卵圆细胞增殖模型,通过原位二步胶原酶灌流法分离到单个肝脏细胞,再经过 Percoll 密度梯度离心分离到肝卵圆细胞。原代大鼠肝细胞/肝卵圆细胞各分为3组:UMSC 组、原代大鼠肝细胞组/肝卵圆细胞组及 UMSC 与原代大鼠肝细胞共培养组/UMSC 与肝卵圆细胞共培养组,分别于第1、3、6、8天通过MTT 法检测各组细胞增殖能力,于第3天通过 ELISA 法检测各组细胞培养上清液中白蛋白含量。结果UMSC 与原代大鼠肝细胞共培养组在第3、6、8天的 A 值均比相应时间点的 UMSC 组 A 值与原代大鼠肝细胞组 A 值之和大(P <0.05);UMSC 与肝卵圆细胞共培养组在第3、6、8、10天的 A 值均比相应时间点的 UMSC 组 A 值与肝卵圆细胞组 A 值之和大(P <0.05)。UMSC 无白蛋白分泌能力,UMSC 与原代大鼠肝细胞共培养组培养上清液中白蛋白水平为(266.21±50.44)ng/mL,较原代大鼠肝细胞组(130.79±22.10)ng/mL 高(P =0.013);UMSC 与肝卵圆细胞共培养组培养上清液中白蛋白水平((49.64±3.56)ng/mL)较肝卵圆细胞组(13.54±1.53)ng/mL 高(P =0.000)。结论UMSC 在体外可以促进原代大鼠肝细胞及肝卵圆细胞的存活和增殖,并可增强其分泌白蛋白的作用。
目的:探討大鼠臍帶間充質榦細胞(UMSC)對原代大鼠肝細胞及肝卵圓細胞增殖與功能的影響。方法採用2-乙酰氨基芴加肝髒三分之二切除術建立肝卵圓細胞增殖模型,通過原位二步膠原酶灌流法分離到單箇肝髒細胞,再經過 Percoll 密度梯度離心分離到肝卵圓細胞。原代大鼠肝細胞/肝卵圓細胞各分為3組:UMSC 組、原代大鼠肝細胞組/肝卵圓細胞組及 UMSC 與原代大鼠肝細胞共培養組/UMSC 與肝卵圓細胞共培養組,分彆于第1、3、6、8天通過MTT 法檢測各組細胞增殖能力,于第3天通過 ELISA 法檢測各組細胞培養上清液中白蛋白含量。結果UMSC 與原代大鼠肝細胞共培養組在第3、6、8天的 A 值均比相應時間點的 UMSC 組 A 值與原代大鼠肝細胞組 A 值之和大(P <0.05);UMSC 與肝卵圓細胞共培養組在第3、6、8、10天的 A 值均比相應時間點的 UMSC 組 A 值與肝卵圓細胞組 A 值之和大(P <0.05)。UMSC 無白蛋白分泌能力,UMSC 與原代大鼠肝細胞共培養組培養上清液中白蛋白水平為(266.21±50.44)ng/mL,較原代大鼠肝細胞組(130.79±22.10)ng/mL 高(P =0.013);UMSC 與肝卵圓細胞共培養組培養上清液中白蛋白水平((49.64±3.56)ng/mL)較肝卵圓細胞組(13.54±1.53)ng/mL 高(P =0.000)。結論UMSC 在體外可以促進原代大鼠肝細胞及肝卵圓細胞的存活和增殖,併可增彊其分泌白蛋白的作用。
목적:탐토대서제대간충질간세포(UMSC)대원대대서간세포급간란원세포증식여공능적영향。방법채용2-을선안기물가간장삼분지이절제술건립간란원세포증식모형,통과원위이보효원매관류법분리도단개간장세포,재경과 Percoll 밀도제도리심분리도간란원세포。원대대서간세포/간란원세포각분위3조:UMSC 조、원대대서간세포조/간란원세포조급 UMSC 여원대대서간세포공배양조/UMSC 여간란원세포공배양조,분별우제1、3、6、8천통과MTT 법검측각조세포증식능력,우제3천통과 ELISA 법검측각조세포배양상청액중백단백함량。결과UMSC 여원대대서간세포공배양조재제3、6、8천적 A 치균비상응시간점적 UMSC 조 A 치여원대대서간세포조 A 치지화대(P <0.05);UMSC 여간란원세포공배양조재제3、6、8、10천적 A 치균비상응시간점적 UMSC 조 A 치여간란원세포조 A 치지화대(P <0.05)。UMSC 무백단백분비능력,UMSC 여원대대서간세포공배양조배양상청액중백단백수평위(266.21±50.44)ng/mL,교원대대서간세포조(130.79±22.10)ng/mL 고(P =0.013);UMSC 여간란원세포공배양조배양상청액중백단백수평((49.64±3.56)ng/mL)교간란원세포조(13.54±1.53)ng/mL 고(P =0.000)。결론UMSC 재체외가이촉진원대대서간세포급간란원세포적존활화증식,병가증강기분비백단백적작용。
Objective To explore the effects of UMSC on proliferation and function of primary rat hepatocytes and hepatic oval cells in vitro . Methods Rat model for hepatic oval cell proliferation was established by 2-acetylaminofluorenne and two third partial hepatectomy (2-AAF/PH).Liver cells were isolated by two steps collagenase perfusion via portal vein in site,and then separated to hepatic oval cells by density gradient centrifugation of Percoll. Primary rat hepatocytes/hepatic oval cells were respectively divided into three groups:UMSC group,primary rat hepatocytes/hepatic oval cells group and UMSC co-culture with primary rat hepatocytes/hepatic oval cells group.Cells proliferation rate of each group was measured by MTT on days 1 ,3,6,8,and concentration of albumin in culture medium was measured by ELISA on day 3.Results OD value of UMSC co-culture with primary rat hepatocytes group was significantly higher than the sum of OD values of UMSC group and primary rat hepatocytes group on days 3,6 and 8 (P <0.05);OD value of UMSC co-culture with hepatic oval cells group was higher than the sum of OD values of UMSC group and hepatic oval cells group on 3,6 and 8(P <0.05).UMSC didn’t secrete albumin,but concentration of albumin in culture medium of UMSC co-culture with primary rat hepatocytes group [(266.21 ±50.44)ng/mL]was higher than that in primary rat hepatocytes group [(130.79 ± 22.10)ng/mL,P =0.0013];Concentration of albumin in culture medium of UMSC co-culture with hepatic oval cells group [(49.64±3.56)ng/mL]was higher than that in hepatic oval cells group [(13.54 ± 1 .53 )ng/mL,P = 0.000 ].Conclusion UMSC can promote proliferation of primary rat hepatocytes and hepatic oval cells in vitro ,and increase secretion of albumin.